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Multipedal DNA Walker Biosensors Based on Catalyzed Hairpin Assembly and Isothermal Strand-Displacement Polymerase Reaction for the Chemiluminescent Detection of Proteins
ACS Sensors ( IF 8.2 ) Pub Date : 2018-06-25 00:00:00 , DOI: 10.1021/acssensors.8b00129 Ningxing Li 1 , Mingyuan Du 1 , Yucheng Liu 1 , Xinghu Ji 1 , Zhike He 1
ACS Sensors ( IF 8.2 ) Pub Date : 2018-06-25 00:00:00 , DOI: 10.1021/acssensors.8b00129 Ningxing Li 1 , Mingyuan Du 1 , Yucheng Liu 1 , Xinghu Ji 1 , Zhike He 1
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In this study, two kinds of sensitive biosensors based on a multipedal DNA walker along a three-dimensional DNA functional magnet particles track for the chemiluminescent detection of streptavidin (SA) are constructed and compared. In the presence of SA, a multipedal DNA walker was constructed by a biotin-modified catalyst as a result of the terminal protection to avoid being digested by exonuclease I. Then, through a toehold-mediated strand exchange, a “leg” of a multipedal DNA walker interacted with a toehold of a catalyzed hairpin assembly (CHA)-H1 coupled with magnetic microparticles (MMPs) and opened its hairpin structure. The newly open stem in CHA-H1 was hybridized with a toehold of biotin-labeled H2. Via the strand displacement process, H2 displaced one “leg” of a multipedal DNA walker, and the other “leg” continued to interact with the neighboring H1 to initiate the next cycle. In order to solve the high background caused by the hybridization between CHA-H1 and H2 without a CHA-catalyst, the other model was designed. The principle of the other model (isothermal strand-displacement polymerase reaction (ISDPR)-DNA walker) was similar to that of the above one. After the terminal protection of SA, a “leg” of a multipedal DNA walker was triggered to open the hairpin of the ISDPR-H1 conjugated with MMPs. Then, the biotin-modified primer hybridized with the newly exposed DNA segment, triggering the polymerization reaction with the assistance of dNTPs/polymerase. As for the extension of the primer, the “leg” of a multipedal DNA walker was displaced so that the other “leg” could trigger the proximal H1 to go onto the next cycle. Due to its lower background and stronger signal, a multipedal DNA walker based on an ISDPR had a lower limit of detection for SA. The limit of detection for SA was 6.5 pM, and for expanding the application of the method, the detections of the folate receptor and thrombin were explored. In addition, these DNA walker methods were applied in complex samples successfully.
中文翻译：
基于催化发夹组装和等温链置换聚合酶反应的多足DNA Walker生物传感器用于蛋白质的化学发光检测。
在这项研究中，构建并比较了两种敏感的生物传感器，它们基于多踏板DNA Walker沿着三维DNA功能磁铁粒子轨道进行化学发光检测，以检测链霉亲和素（SA）。在存在SA的情况下，由于末端保护而避免了被核酸外切酶I消化，因此由生物素修饰的催化剂构建了多足的DNA Walker。然后，通过脚趾介导的链交换，形成了多足的“腿”。 DNA Walker与催化发夹组件（CHA）-H1的脚尖相互作用，并与磁性微粒（MMP）耦合，并打开了其发夹结构。CHA-H1中新近开放的茎与生物素标记的H2的前额杂交。通过链置换过程，H2置换了多踏板DNA Walker的一个“腿”，另一个“腿”继续与相邻的H1交互以启动下一个周期。为了解决由CHA-H1和H2在没有CHA催化剂的情况下杂交引起的高背景，设计了另一种模型。其他模型的原理（等温链置换聚合酶反应（ISDPR）-DNA Walker）与上述模型相似。在SA的末端保护之后，触发了多足DNA Walker的“腿”以打开与MMP结合的ISDPR-H1的发夹。然后，生物素修饰的引物与新暴露的DNA片段杂交，在dNTPs /聚合酶的辅助下引发聚合反应。至于引物的延伸，多脚踏DNA Walker的“腿”被置换，以便另一个“腿”可以触发近端H1进入下一个循环。由于其较低的背景和较强的信号，基于ISDPR的多踏板DNA Walker对SA的检测下限较低。SA的检出限为6.5 pM，为扩大该方法的应用范围，探索了叶酸受体和凝血酶的检测方法。此外，这些DNA Walker方法已成功应用于复杂样品中。
更新日期：2018-06-25
中文翻译：
基于催化发夹组装和等温链置换聚合酶反应的多足DNA Walker生物传感器用于蛋白质的化学发光检测。
在这项研究中，构建并比较了两种敏感的生物传感器，它们基于多踏板DNA Walker沿着三维DNA功能磁铁粒子轨道进行化学发光检测，以检测链霉亲和素（SA）。在存在SA的情况下，由于末端保护而避免了被核酸外切酶I消化，因此由生物素修饰的催化剂构建了多足的DNA Walker。然后，通过脚趾介导的链交换，形成了多足的“腿”。 DNA Walker与催化发夹组件（CHA）-H1的脚尖相互作用，并与磁性微粒（MMP）耦合，并打开了其发夹结构。CHA-H1中新近开放的茎与生物素标记的H2的前额杂交。通过链置换过程，H2置换了多踏板DNA Walker的一个“腿”，另一个“腿”继续与相邻的H1交互以启动下一个周期。为了解决由CHA-H1和H2在没有CHA催化剂的情况下杂交引起的高背景，设计了另一种模型。其他模型的原理（等温链置换聚合酶反应（ISDPR）-DNA Walker）与上述模型相似。在SA的末端保护之后，触发了多足DNA Walker的“腿”以打开与MMP结合的ISDPR-H1的发夹。然后，生物素修饰的引物与新暴露的DNA片段杂交，在dNTPs /聚合酶的辅助下引发聚合反应。至于引物的延伸，多脚踏DNA Walker的“腿”被置换，以便另一个“腿”可以触发近端H1进入下一个循环。由于其较低的背景和较强的信号，基于ISDPR的多踏板DNA Walker对SA的检测下限较低。SA的检出限为6.5 pM，为扩大该方法的应用范围，探索了叶酸受体和凝血酶的检测方法。此外，这些DNA Walker方法已成功应用于复杂样品中。
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